Dulbecco smodified eagle medium dmem

RAW264.7
cells were obtained from the Korean Cell Line Bank. Dulbecco’s
Modified Eagle Medium (DMEM), penicillin-streptomycin, and fetal bovine
serum (FBS) were obtained from GenDEPOT. Silymarin, dimethyl sulfoxide
(DMSO), soluble 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium
bromide (MTT), Sodium selenite, and lipopolysaccharide (LPS) were
purchased from Sigma-Aldrich. Sodium selenite was purchased from Sigma
Chemicals Co. The 740Y–P was purchased from MedChemExpress
(MCE). Primary antibodies against PI3K, p-PI3K, AKT, and p-AKT were
obtained from Abcam, and antibodies against IκBα, p-IκBα,
NF-κB, p-NF-κB, and β-actin were purchased from
Cell Signaling Technology.

The striatal neuronal culture method was described by Lee et al. [22 (link)]. Briefly, striatal tissue was dissected from Sprague–Dawley rat pups (E19~E20) and enzymatically digested into a single-cell suspension with a mild protease solution (0.25% trypsin-EDTA; Gibco, Grand Island, NY, USA) at 37℃ for 30 min. After removal of the digestion solution, the tissue was washed three times in dissociation media containing 90 mM of Na₂SO₄, 30 mM of K₂SO₄, 16 mM of MgCl₂, 0.25 mM of CaCl₂, 32 mM of HEPES, and 0.01% phenol red (Sigma, St. Louis, MO, USA), with a pH of 7.7, and transferred to a tissue-culture medium (Dulbecco's modified Eagle medium [DMEM]; GenDEPOT, Barker, TX, USA) containing 10% fetal bovine serum, 2% B27 (Gibco), and 100 U/ml of penicillin/streptomycin. Next, the tissue was triturated into a single-cell suspension via a 5-ml serological pipette and plated at a density of 2.2×10⁵ cells/cm² on poly-L-lysine-coated 24-well plates.